鸡杆菌中国分离株RAPD分型方法的建立与应用  

Development and application of random amplified polymorphic DNA as a typing method for Gallibacterium anatis isolated in China

作　　者：陈陆[1] 付仁一[1] 杨霞[1] 王新卫[1] 皇甫和平[1] 徐雪[1] 刘慧敏[1] 郑鹿平[1] 郭伦涛[1] 彭民泽 王川庆[1] 

机构地区：[1]河南农业大学禽病研究所,河南郑州450002 [2]三门峡市农业科学研究院,河南三门峡472000

出　　处：《中国兽医学报》2011年第4期465-469,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30972187)

摘　　要：对9株鸡杆菌进行了血清分型,并应用8条随机引物对该9株鸡杆菌和3个参考菌株(1株巴氏杆菌、1株大肠杆菌和1株链球菌)共12株细菌做随机扩增多态性DNA(RAPD)分型研究。结果显示,9株鸡杆菌分别属于血清Ⅰ型(1/9)、血清Ⅱ型(3/9)和血清Ⅳ型(5/9)。所用8条随机引物中有3条呈现良好的多态性和稳定性,可产生差异显著的指纹图谱。采用SPSS13.0软件分析了不同菌株间的遗传距离,并依此绘制出菌株间的亲缘关系树状图。12株细菌共分为4个聚类群,其中9株鸡杆菌位于同一聚类群中,且又可分为4个聚类亚群。3个参考菌株各自形成一个独立的聚类群。不同血清型的鸡杆菌菌株具有相似的指纹图,同一血清型的菌株指纹图存在差异。结果表明,RAPD基因分型是目前鸡杆菌分子流行病学调查及病原分离鉴定比较理想的方法之一。In order to provide the reference for investigating the epidemic origin of Gallibacterium anatis,typing of Gallibacterium anatis and the relationship between random amplified polymorphic DNA（RAPD） typing and serotyping of Gallibacterium anatis isolations.nine strains of Gallibacterium anatis isolated from layer flocks in Henan were systematically classified by serotyping and genotyping using randomly amplified polymorphic DNA（RAPD） with eight random primers,and three reference strains of Pasteurella multocida,Escherichia coli and Streptococcus suis were used as control.The Gallibacterium anatis infection was mainly caused by serotype Ⅰ（1/9）,serotype Ⅱ（3/9） and serotype Ⅳ（5/9）.In RAPD study,the results showed that three efficient primers were screened from eight random primers.The heredity distance of different strains was got by SPSS13.0 and the polygenetic tree among these strains was drawn.According to the heredity distance,eleven strains used in this study can be divided into 4 c1ustering groups,and nine strains of Gallibacterium anatis were divided into 4 c1ustering subgroups,three reference strains formed three c1ustering groups,respectively.Gallibacterium anatis isolates of different serotype had similar finger print map,Gallibacterium anatis isolates of the same serotype did not have the same finger print map.According to it,RAPD typing is one of better methods in molecular epidemiological investigation,isolation and identification of Gallibacterium anatis,at present.

关 键 词：鸡杆菌 血清分型 RAPD 基因分型 

分 类 号：S852.61[农业科学—基础兽医学]